Document Detail


Labeling D+ RBCs for flow cytometric quantification of fetomaternal hemorrhage after the RBCs have been coated with anti-D.
MedLine Citation:
PMID:  11493739     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: D- patients may receive Rh immunoglobulin (RhIg) before blood samples are taken for estimation of the volume of fetomaternal hemorrhage (FMH) by flow cytometry. Anti-D bound to the fetal D+ cells may then block the binding of conjugated D MoAb. This may reduce the fluorescence of the D+ cells, which would lead to ambiguity over setting the positions of the markers on histograms and may result in erroneous values of FMH. STUDY DESIGN AND METHODS: Labeling methods were compared by using FITC-BRAD 3 (anti-D) and/or FITC-anti-IgG (Fab fragment) with mixtures of D+ (R1r) and D- (rr) cells when the D+ cells had first been coated with various amounts (0 molecules/cell and 600-13,000 molecules/cell) of anti-D (RhIg). Variables examined were antibody concentrations, the order and times of incubation with the antibodies, and the effect of washing between the uptake of the antibodies used. RESULTS: In all cases, D+ cells were strongly labeled after incubation with 50 microL of FITC-BRAD-3 and then after washing with 50 microL of FITC-anti-human IgG, with both incubations being for 30 minutes at 37 degrees C. With this double-staining procedure, the fluorescence of D+ cells was found to be similar regardless of how much anti-D (RhIg) was previously bound and greater than that with FITC-BRAD-3 alone, giving an enhanced signal-to-noise ratio. CONCLUSION: As the testing laboratory may not know if the patient has received prophylactic RhIg, this labeling method would be suitable for all samples.
Authors:
B M Kumpel
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Publication Detail:
Type:  Comparative Study; Journal Article    
Journal Detail:
Title:  Transfusion     Volume:  41     ISSN:  0041-1132     ISO Abbreviation:  Transfusion     Publication Date:  2001 Aug 
Date Detail:
Created Date:  2001-08-08     Completed Date:  2001-08-23     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0417360     Medline TA:  Transfusion     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1059-63     Citation Subset:  IM    
Affiliation:
International Blood Group Reference Laboratory, Southmead Road, Bristol, United Kingdom. belinda.kumpel@nbs.nhs.uk
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MeSH Terms
Descriptor/Qualifier:
Adult
Antibodies, Monoclonal / diagnostic use
Erythrocytes / immunology*,  metabolism
Female
Fetal Blood
Fetomaternal Transfusion / blood,  diagnosis*,  immunology
Flow Cytometry / methods*,  standards
Humans
Isoantibodies / analysis,  metabolism
Pregnancy
Rh-Hr Blood-Group System / immunology*
Sensitivity and Specificity
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Isoantibodies; 0/Rh-Hr Blood-Group System; 0/Rho(D) antigen

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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